Production of proteins using transposon-based vectors

摘要

Novel compositions for the in vitro or in vivo production of specific proteins are provided. The compositions comprise components of vectors, such as a vector backbone, a promoter, and a gene of interest that encodes for the protein of interest, and the transposon-based vectors comprising these components. Also provided are methods of making these compositions and methods of using these compositions for the production of desired proteins in vivo or in transfected cells in vitro.

主权项

1. A vector comprising:
a modified transposase gene operably-linked to a first promoter, wherein the nucleotide sequence 3′ to the first promoter comprises a Kozak sequence, and wherein a plurality of the first twenty codons of the modified transposase gene are modified from the wild-type sequence by changing the nucleotide at the third base position of the codon to an adenine or thymine without modifying the amino acid encoded by the codon; one or more genes of interest operably-linked to one or more additional promoters, wherein the one or more genes of interest encodes a protein of interest selected from the group consisting of erythropoietin, bone morphogenic protein, high density lipoprotein, platelet derived growth factor, enterokinase, vascular endothelial cell derived growth factor, luteinizing hormone, trastuzumab, etanercept, colony stimulating factor, and human chorionic gonadotropin, and wherein the one or more genes of interest and their one or more operably-linked promoters are flanked by transposase insertion sequences recognized by a transposase encoded by the modified transposase gene; and one or more insulator elements located between the transposase insertion sequences and the one or more genes of interest, wherein the one or more insulator elements comprise a lysozyme replicator element, a combination of a lysozyme replicator element and an HS4element, or a matrix attachment region element, wherein the start codon of the gene of interest is located about 2502 bp from the one or more insulator elements located 5′ to the start codon of the gene of interest.